Epidermal Growth Factor Composition, A Process Therefor and Its Application

ABSTRACT

A composition for treating a wound, wherein the composition can comprise therapeutically effective amount of an epidermal growth factor and a physiologically acceptable agent, wherein the physiologically acceptable agent comprises at least one of a stabilizer, a preservative, a thickening agent, carrier/diluent, and optionally pH regulating agent and humectant.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.11/915,727, filed Jun. 16, 2008, which is the National Stage ofInternational Application No. PCT/IN2006/000168, filed May 18, 2006, andclaims priority to Indian Patent Application No. 00642CHE2005, filed May27, 2005, which are incorporated by reference herein in their entirety.

FIELD OF THE INVENTION

The invention particularly relates to a stable composition of epidermalGrowth Factor (herein referred to as EGF). Particularly, the inventionrelates to a stable EGF composition involving novel recombinant EGF(rEGF). More particularly, the invention relates to a compositioncomprising an epidermal growth factor and a physiologically acceptableagent, wherein the physiologically acceptable comprises at least one ofa stabilizer, a preservative and a thickening agent. Further theinvention also relates to the application/use of the composition fortreating wounds. The invention also describes the process forpreparation of the said stable composition having rEGF astherapeutically active ingredient.

BACKGROUND OF THE INVENTION

Epidermal Growth Factor (EGF) is a single chain polypeptide havingmolecular weight of approximately 62 kDa and comprised of 53 amino acidresidues including three disulphide bonds. Epidermal Growth Factor (EGF)is one of the most potent, biologically available entities that play avery important role in wound healing. It binds specifically to the EGFreceptors present on the cell surface, thus triggering complexbiochemical mechanisms in the cells to trigger the growth. EGF isbiologically unstable and physiologically nonhomogenous, particularly inthe presence of moisture. EGF has a short life span of about an hourwhereas DNA synthesis at the site of the wound requires about eight totwelve hours. EGF exhibits loss of biological activity due todenaturation, decomposition, condensation and precipitation due toproteolytic enzymes. This is disadvantageous because such loss ofactivity makes it impractical to store aqueous preparations of epidermalgrowth factor for extended periods of time. The use of EGF formulationsfor wound healing is known in the art. To overcome this problem and toprovide desired effective wound healing, it is reported that EGF has tobe continuously applied in initial stages of healing. As a result, manyformulations have been developed to increase the stability of EGF.

The prior art known to the inventors include CN 1515315 that relates toa stable synergistic composition containing EGF and bletilla extract.

U.S. Pat. No. 4,944,948 suggests a liposome gel formulation for thedelivery of hEGF to the wound site to overcome the above-mentionedproblems.

EP 0312208 discloses using various water-soluble or water-swellablecarriers for slow release formulations for EGF. This enables release ofEGF for 12 hours or more. However, these formulations are unsuitable forindustrial application due to poor shelf life.

U.S. Pat. No. 4,717,717 advocates using cellulose derivative togetherwith EGF to enhance stabilization.

EP 0312208 ('208) discloses aqueous gel formulation of EGF forcontrolled delivery of the active ingredient employing various watersoluble polymer as a base for providing viscosity ranging from 1000 to12,000,000 cps at room temperature.

U.S. Pat. No. 4,944,948 describes gel formulation of EGF using neutralphospholipids, negatively-charged phospholipids, and cholesterol.

Though these formulations are capable of continuously releasing EGF for12 hours or more they are unsuitable for industrial manufacture beingunstable in long term storage. Further, due to high viscosity of theformulation disclosed in '208 it forms barrier for migration ofepidermal cells. Additionally, it poses problems in application atdelicate wound sites.

U.S. Pat. No. 5,130,298 and EP publication No. 398615 teaches mixing EGFwith metal cation for preventing degradation of EGF through ionicbinding. This increases the stability of EGF to about 2 months at 4° C.This formulation also proves to be unsuitable for industrialapplication.

US 2003050238 advises using acidic polymer such as carboxyvinyl polymeras a base to solve the above-mentioned problems of shelf life andunsuitability for utilizing in industrial fields. The patent claims tohave increased the shelf life.

PCT Application No. WO99/44631 attempts to extend shelf life of hEGF.However, none of these solutions were found successful and the stabilityof the enzyme remains a problem as far as it stays in aqueousenvironment above 0° C.

It is also noticed that EGF degrades over time to form multiple speciesof the EGF molecule, which are believed to be degradation products. Suchdegradation occurs naturally as a result of environmental factors suchas light, which can cause photo-oxidation; changes in pH; changes inionic strength; changes in temperature; and physical manipulation of themolecule. This reduces the shelf life of an EGF formulation overextended storage.

Hence, there is an imperative need to develop a stable formulation ofEGF. The inventors after conducting considerable research have developedEGF composition having stability over two years while maintaining itsefficacy. Further, it provides public with useful choice. Afterprolonged research it has been found out that the compositions of EGFparticularly rEGF, when prepared using physiologically acceptable agentscomprising charged protein stabilizers specifically in combination withpreservative, thickening agent, pH regulating agent and a carrier provesto be stable for about 2 years.

SUMMARY OF THE INVENTION

The invention relates to a composition for treating a wound wherein thecomposition can comprise therapeutically effective amount of anepidermal growth factor and a physiologically acceptable agent, whereinthe physiologically acceptable agent comprises at least one of astabilizer, a preservative, a thickening agent, carrier/diluent, andoptionally pH regulating agent and humectant. The stabilizer can be oneof L-lysine Hydrochloride, Mannitol and carboxymethyl cellulose, thepreservative can be a salt of p-hydroxy benzoic acid and the thickeningagent can be polyacrylic acid. Triethanol amine being used mainly as apH regulating agent also serves the purpose of crosslinking Humectantcan be glycerol. The physiologically acceptable agent can furtherinclude alpha linolenic acid.

According to an embodiment, the composition can further comprisetherapeutically effective amount of a therapeutic agent. The therapeuticagent is selected from a group consisting of a synthetic anti-infectiveagent, a recombinant biological active ingredient, a recombinantantibiotic, a natural product, protein free blood extract, an immunemodulator, clobetasol propiate, and, a vaso constrictor.

According to another embodiment, a method for treating a wound cancomprise a step of administering a composition to the wound, wherein thecomposition comprises therapeutically effective amount of an epidermalgrowth factor (EGF) and a physiologically acceptable agent, wherein thephysiologically acceptable agent comprises at least one of a stabilizer,a preservative and a thickening agent.

According to another embodiment, a process of making a composition fortreating a wound comprises dissolving therapeutically effective amountof a physiologically acceptable agent in water to obtain a mixture,wherein the physiologically acceptable agent comprises at least one of astabilizer, a preservative and a thickening agent; and, addingtherapeutically effective amount of an epidermal growth factor to themixture to obtain the composition.

The composition of this invention can be formulated as semi solid formsuch as gel, cream, ointment for topical application, or suitable forparentaral or injectable administration by appropriately adjusting thequantities of respective responsible ingredients.

DETAILED DESCRIPTION OF THE INVENTION

As required, detailed embodiments of the present invention are disclosedherein; however, it is to be understood that the disclosed embodimentsare merely exemplary of the invention, which can be embodied in variousforms. Therefore, specific structural and functional details disclosedherein are not to be interpreted as limiting, but merely as a basis forthe claims and as a representative basis for teaching one skilled in theart to variously employ the present invention in virtually anyappropriately detailed structure. Further, the terms and phrases usedherein are not intended to be limiting but rather to provide anunderstandable description of the invention.

The terms “a” or “an”, as used herein, are defined as one or more thanone. The term “plurality”, as used herein, is defined as two or morethan two. The term “another”, as used herein, is defined as at least asecond or more. The terms “including” and/or “having”, as used herein,are defined as comprising (i.e., open language).

The present invention relates to a composition for treating a wound.According to one embodiment, the composition comprises therapeuticallyeffective amounts of an epidermal growth factor and a physiologicallyacceptable agent, wherein the physiologically acceptable agent comprisesat least one of a stabilizer, a preservative a thickening agent,carrier/diluent, and optionally pH regulating agent and humectant.According to an embodiment, the epidermal growth factor can be arecombinant epidermal growth factor. The epidermal growth factor can bein the range of about 0.001% to about 0.1% (w/w).

According to an embodiment, the stabilizer comprises one of L-lysineHydrochloric acid, mannitol and carboxymethylcellulose, the preservativecomprises a salt of p-hydroxy benzoic acid and the thickening agentcomprises polyacrylic acid. According to a further embodiment, thepreservative comprises sodium methyl paraben and sodium propyl paraben.The physiologically acceptable agent also includes components, inaddition to the epidermal growth factor, which are suitable foradministration to the patient being treated in accordance with theinvention. For example, the physiologically acceptable agent can includecomponents, which result in a stable composition or increase theefficacy of the composition. According to another embodiment, thephysiologically acceptable agent further comprise water,triethanolamine, alpha linolenic acid, and glycerol. The mannitol can bein the range of about 0.5% to about 10% w/w. The L-lysine hydrochloricacid can be in the range of about 0.1% to about 2% w/w. The polyacrylicacid can be in the range of about 0.25% to about 3% w/w. Thetriethanolamine can be in the range of about 1% to about 20% (w/w). Thesodium methyl paraben can be about 0.016% to about 0.18% (w/w) and thesodium propyl paraben can be about 0.01% to about 0.02% (w/w). Theglycerol can be about 1.0% to about 2.5% (w/w) and thecarboxymethylcellulose can be in the range of about 0.6% to about 1.6%(w/w). The water can be in the range of about 50% to about 95% (w/v).

According to another embodiment, the composition can further comprisewater of about 98% (w/v), wherein the polyacrylic acid is in the rangeof about 0.25% to about 0.8% (w/w), triethanolamine is in the range ofabout 1% to about 20% (v/v), sodium methyl paraben is about 0.18% w/wand sodium propyl paraben is about 0.02% (w/w). The composition can beformulated as a semi-solid drug delivery formulation.

According to another embodiment, the composition can further comprisewater of about 97.5% to about 99% w/v, wherein thecarboxymethylcellulose is in the range of about 0.6% to about 1.6%(w/w). The composition can be formulated as a capsule.

According to another embodiment, the composition can further comprisetherapeutically effective amount of a therapeutic agent. According to anembodiment, the therapeutic agent can include components, which canassist in the wound-healing process and are suitable for administrationto patients. The therapeutic agent can also include components, whichperform additional functions such as increasing the uptake of oxygen,moisturizing or increasing the proliferation of granulocytes ormacrophages, etc. The therapeutic agent can include components whichperform functions in addition to those performed by the epidermal growthfactor. The therapeutic agent can be selected from a group consisting ofa synthetic anti-infective agent, a recombinant biological activeingredient, a recombinant antibiotic, a natural product, protein freeblood extract, an immune modulator, clobetasol propiate, and, a vasoconstrictor.

According to an embodiment, the synthetic anti-infective agent may beselected from a synthetic group comprising erythromycin, mupricin,soframycin, clindamycin phosphate, fluconazole and silver sulphadiazine.The synthetic anti-infective agent can be in the range of about 0.1% toabout 5% (w/w). According to another embodiment, the syntheticanti-infective agent can comprise silver sulphadiazine in the range ofabout 0.05% to about 0.5% (w/w).

According to an embodiment, the biological active can be selected from abiological group consisting of hyaluronic acid, a granulocyte colonystimulating factor, and a transforming growth factor-alpha. Hyaluronicacid (HA) is found extensively in nature in micro organisms, in humansand also in animals. It is highly viscous lubricant and hence can beused as a moisturizer. According to an embodiment, hyaluronic acid canbe used in combination with EGF for reendothelialization in the anteriorchamber of the eye. The hyaluronic acid can be in the range of about0.1% to about 5% (w/v). The Granulocyte macrophage-colony stimulatingfactor (GM-CSF) increases the proliferation of granulocytes andmacrophages. The granulocyte colony stimulating factor can be in therange of about 0.002% to about 0.004% (w/w). The Transforming GrowthFactor—Alpha promotes normal wound healing through a concerted effortwith Epidermal Growth Factor and Platelet-Derived Growth Factor (PDGF).The transforming growth factor-alpha can be in the range of about 0.001%to about 0.1% (w/w).

According to an embodiment, the recombinant antibiotic compriseslysostaphin. Lysostaphin is a biological antibiotic produced byStaphylococcus. Lysostaphin can be cloned and expressed in E. Coli, andpurified and formulated at suitable concentration to give antibacterialeffects in combination with EGF at therapeutically effectiveconcentration. The lysostaphin can be in the range of about 0.001% toabout 0.1% (w/w).

According to another embodiment, the natural product can be selectedfrom the group which can comprise aloe vera, honey, turmeric and sandalwood. Turmeric is an excellent natural antibiotic. The aloe vera can bein the range of about 1% to about 2% (w/w), the honey can be in therange of about 0.1% to about 1% (w/w), the turmeric can be in the rangeof about 5% to about 80% (w/w) and the sandal wood can be in the rangeof about 0.2% to about 1% (w/w).

According to another embodiment, the composition can comprise theprotein free blood extract in the range of about 0.02% (w/w). Proteinfree blood extract improves the utilization of oxygen and promotes theuptake of nutrients into the cell.

According to an embodiment, the immune modulator comprisesbeta-1,3-D-glucan. The composition can further comprise an effectiveamount of vitamin A, vitamin C and vitamin D, wherein the immunemodulator comprises beta-1,3-D-glucan in the range of about 0.5% toabout 2% (w/w). Beta-1,3-D-glucan itself can elicit broad anti-infectiveeffects such as antibody response against Staphylococcus aureus,Escherichia coil, Candida albicans, Pneumocytis carinii, Listeriamonococytogenesis, Leishmania donovani, and Herpes simplex.

According to an embodiment, the vaso constrictor compriseshydrocortisone acetate. The composition can further comprise lidocainein the range of about 2% to about 5% (w/w), zinc oxide in the range ofabout 2% to about 5% (w/w), allantoin in the range of about 0.25% toabout 2.5% (w/w), wherein the vaso constrictor comprises hydrocortisoneacetate in the range of about 0.15% to about 0.25% (w/w). According toan embodiment, the composition can be used in treating hemorrhoids andfissure proctitis. The composition can also provide symptomatic painrelief.

According to an embodiment, the composition can further comprise zincoxide in the range of about 2% to about 5% (w/w) and salicylic acid inthe range of about 0.1% to about 5% (w/w), wherein the therapeutic agentcomprises clobetasol propionate in the range of about 0.1% to about 1%(w/w).

According to another embodiment, the composition can be formulated as atopical formulation. The topical formulation is selected from a groupcomprising gels, sprays, ointments, creams and lotions. According to anembodiment, the composition can also be as an oral formulation suitablefor oral administration and a parenteral formulation suitable forparenteral administration. The formulations for oral administration cancomprise capsules. The formulations for parenteral administration caninclude injections. However, the scope of the invention is not justlimited to these formulation types but can also be expanded to otherformulation types known to those skilled in the art.

According to another embodiment, a method for treating a wound cancomprise a step of administering an effective amount of the composition,wherein the composition comprises therapeutically effective amount of anepidermal growth factor and a physiologically acceptable agent, whereinthe physiologically acceptable agent comprises at least one of astabilizer, a preservative, a thickening agent, carrier/diluent, andoptionally pH regulating agent and humectant. According to anembodiment, the stabilizer comprises one of L-lysine Hydrochloric acid,mannitol, and carboxymethylcellulose, the preservative comprises a saltof p-hydroxy benzoic acid and the thickening agent comprises polyacrylicacid. According to a further embodiment, the preservative comprisessodium methyl paraben and sodium propyl paraben. According to anotherembodiment, the physiologically acceptable agent can further comprisewater as carrier, triethanolamine, alpha linolenic acid, and glycerol(humectant).

According to another embodiment, the composition can further comprisetherapeutically effective amounts of a therapeutic agent. Thetherapeutic agent can be selected from a group comprising a syntheticanti-infective agent. a recombinant biological active ingredient, arecombinant antibiotic, a natural product, protein free blood extract,an immune modulator, clobetasol propiate, silver sulphadiazine and avaso constrictor.

According to another embodiment, the composition can be used in treatingwounds such as ulcers, for example, diabetic foot ulcer, corneal ulcer,gastric ulcer, venous ulcer, arterial ulcer and pressure ulcer, skinburns caused by irradiation used in cancer therapy, sports injury,thermal injury, chemical injury, physical injury, osteomyelitis,dermatitis, muscle soreness, joint soreness, muscle stiffness, jointstiffness, laceration, scarring, surgical wound, bedsores and cuts dueto environmental factors, marks due to childbirth, loss of sensation dueto Mycobacterium leprae infection or by infections which cause similarconditions, hemorrhoids and gastro duodenal ulcer, growth of hairfollicles, and in all other skin ailments which need to be repaired.

According to another embodiment, a process of preparing a compositionfor treating a wound is provided. The process can comprise initiallydissolving therapeutically effective amount of a physiologicallyacceptable agent in water to obtain a mixture, wherein thephysiologically acceptable agent comprises at least one of a stabilizer,a preservative and a thickening agent. According to an embodiment, thestabilizer comprises one of L-lysine Hydrochloric acid andcarboxymethylcellulose, the preservative comprises a salt of p-hydroxybenzoic acid and the thickening agent comprises polyacrylic acid.According to a further embodiment, the preservative comprises sodiummethyl paraben and sodium propyl paraben. The process can furthercomprise adding therapeutically effective amount of an epidermal growthfactor to the mixture to obtain the composition for treating the wound.

According to another embodiment, the process can further comprise addingtherapeutically effective amounts of at least one of L-lysinehydrochloric acid, mannitol and polyacrylic acid to the mixture toobtain a suspension. A therapeutically effective amount of glycerol canbe added to this suspension. The suspension can be stirred so as toallow it to swell. The pH of the suspension can be maintained at about6.3 to 6.4. The pH of the suspension can be maintained by the additionof triethanolamine to the suspension.

According to another embodiment, the process can also further compriseadding therapeutically effective amount of a therapeutic agent to thecomposition. The therapeutic agent can be selected from a groupcomprising a synthetic anti-infective agent, a biological active, arecombinant antibiotic, a natural product, protein free blood extract,an immune modulator, clobetasol propiate and a vaso constrictor. Thesynthetic anti-infective agent can be selected from a synthetic groupcomprising erythromycin, mupricin, soframycin, clindamycin phosphate,fluconazole and silver sulphadiazine, the biological active can beselected from a biological group consisting of hyaluronic acid, agranulocyte colony stimulating factor, and a transforming growthfactor-alpha, the recombinant antibiotic comprises lysostaphin, thenatural product can be selected from the group consisting aloe vera,honey, turmeric and sandal wood, the immune modulator comprisesbeta-1,3-D-glucan, and the vaso constrictor comprises hydrocortisoneacetate.

The composition for treating a wound is described with reference to thefollowing examples. Percentages in the examples are stated as apercentage of the total composition. These examples are provided as anillustration of the invention and are not intended to limit the scope ofthe invention.

Example 1

A mixture was obtained by dissolving about 0.165% of sodium methylparaben and about 0.018% sodium propyl paraben in purified water ofabout 91.57%. Subsequently, about 0.458% of L-lysine hydrochloride,about 4.579% of mannitol and about 0.916% polyacrylic acid were added tothe mixture to obtain a suspension. The suspension was further allowedto swell by stirring. An effective amount of glycerol of about 2.289%was added to this suspension. The pH was maintained at about 6.3 to 6.4with the addition of effective amount of triethanolamine. Finally, atherapeutically effective amount of EGF of about 0.005% was added toobtain the composition for treating the wound.

Example 2

A mixture was obtained by dissolving about 0.162% of sodium methylparaben and about 0.018% sodium propyl paraben in purified water ofabout 89.916%. Subsequently, about 0.450% of L-lysine hydrochloride,about 4.496% of mannitol and about 2.697% polyacrylic acid were added tothe mixture to obtain a suspension. The suspension was further allowedto swell by stirring. An effective amount of glycerol of about 2.248%was added to this suspension. The pH was maintained at about 6.3 to 6.4with the addition of effective amount of triethanolamine. Finally, atherapeutically effective amount of EGF of about 0.013% was added toobtain the composition for treating the wound.

Example 3

A mixture was obtained by dissolving about 0.157% of sodium methylparaben and about 0.017% sodium propyl paraben in purified water ofabout 87.497%. Subsequently, about 0.437% of L-lysine hydrochloride,about 8.750% of mannitol and about 0.875% polyacrylic acid were added tothe mixture to obtain a suspension. The suspension was further allowedto swell by stirring. An effective amount of glycerol of about 2.187%was added to this suspension. The pH was maintained at about 6.3 to 6.4with the addition of effective amount of triethanolamine. Finally atherapeutically effective amount of EGF of about 0.079% was added toobtain the composition for treating the wound.

Various combinations of EGF were thus carried out and the combinationswere subjected to thermal stability as per ICH guidelines. Pre-clinicalstudies for safety of the formulations with varying percentages of EGF,were conducted in both rats and rabbits, as per the approved guidelines.Briefly, acute toxicity studies were carried out for 8 days, sub-acutestudies were carried on for 30 days and ocular irritation study wasconducted for 7 days.

Results indicated that the animals were normal during the study and noneof the animals showed any signs of toxicity related to general behavior,central and autonomic nervous system, respiratory and circulatorysystem. The wound region was devoid of bleeding and erythema through theperiod of study.

The growth rate and food consumption in control and treated animals werenormal & did not differ significantly. The biochemical parametersstudied in control and treated groups were within the normal range andno abnormality was observed. The statistical evaluation revealed thatthe differences observed between the control and the different treatmentgroups were not statistically indicating that rh EGF has not influencedany of the parameters studied. There were no observable changes in thehematological parameters between the control and treated groups.

There was no observable anti-body response in the treated groups withEGF, in both rats and rabbits. The protein content in the control andthe treated groups did not differ significantly in both the speciesstudied. The collagen content was significantly increased in medium andhigh dosage groups in males and females of both the species on the 15thand the 31st days.

Subsequently, the formulation was also tested in patients suffering fromdonor site skin grafts, burns and diabetic foot ulcers.

Data revealed that the healing time for donor site skin graft in thetest group was 7 days in comparison to the control, which was 13 days(P<0.001). In treatment with burns cases, the healing time was 9 days inthe test group while in the control it was observed to be 20 days(P<0.05).

The healing rate in diabetic foot was compared at 7 weeks and 15 weeks.It was found that by 7 weeks, 6.25% of the control cases were healed,while in the treated group 56% cases were healed. The value of P<0.001states the above significance. It was observed that by 15 weeks, 37.5%of the control cases were healed, while in the test group 88% cases werehealed. The value of P<0.005 states the above significance.

The stability of EGF in the gel form has been conducted in both realtime (5+/−3° C.) and accelerated time (25+/−2 and 60% RH+/−5%). Thestability of the formulation was monitored for 2 years at real time andfor 6 months at accelerated time. Data indicated that the molecule wasstable and showed no significant drop in activity during the period ofstudy.

The present study was carried out in wounds, which are more than 20cms², and about 10-40 mm in depth.

The scope of the combination is not limited just by the above range, butalso to other combinations, which can still maintain the same functionalactivity of EGF known to those who are skilled in the art. Thiscombination is used as curative for skin grafts, burns, cuts, diabeticfoot ulcer, post operative wounds, bed sores, vitiligo and repair ofskin damage caused by irradiation leading to cancer.

Example 4

A mixture was obtained by dissolving sodium methyl paraben of about0.163% and sodium propyl paraben of about 0.018% in purified water ofabout 90.739%. Subsequently about 0.454% of L-lysine hydrochloride,about 4.537% of mannitol and about 0.907% of polyacrylic acid were addedto the mixture to obtain a suspension. The suspension was furtherallowed to swell by stirring. An effective amount of glycerol of about2.268% was added to the suspension. The pH was maintained at about 6.3to 6.4 with the addition of effective amount of triethanolamine. Finallytherapeutically effective amount of the antibiotic infective agent ofabout 0.907% and the epidermal growth factor (EGF) about 0.005% wasadded to obtain the composition for treating the wound.

Example 5

A mixture was obtained by dissolving sodium methyl paraben of about0.016% and sodium propyl paraben of about 0.018% in purified water ofabout 90.803%. Subsequently about 0.453% of L-lysine hydrochloride,about 4.540% of mannitol and about 0.908% of polyacrylic acid were addedto the mixture to obtain a suspension. The suspension was furtherallowed to swell by stirring. An effective amount of glycerol of about2.270% was added to the suspension. The pH was maintained at about 6.3to 6.4 with the addition of effective amount of triethanolamine. Finallytherapeutically effective amount of the antibiotic infective agent ofabout 0.908% and the epidermal growth factor (EGF) about 0.082% wereadded to obtain the composition for treating the wound.

Example 6

A mixture was obtained by dissolving about 0.164% sodium methyl parabenand about 0.018% sodium propyl paraben in purified water of about91.478%. Subsequently L-lysine hydrochloride of about 0.457%, mannitolof about 4.574% and polyacrylic acid of about 0.914% were added to themixture to obtain a suspension. The suspension was further allowed toswell by stirring. To this suspension, effective amount of glycerol ofabout 2.286% was added. The pH was maintained at about 6.3 to 6.4 withthe addition of effective amount of triethanolamine. Finallytherapeutically effective amount of hyaluronic acid of about 0.091%,followed by therapeutically effective amount of the epidermal growthfactor (EGF) of about 0.014% was added to obtain the composition fortreating the wound.

Example 7

A mixture was obtained by dissolving about 0.158% sodium methyl parabenand about 0.017% sodium propyl paraben in purified water of about88.269%. Subsequently L-lysine hydrochloride of about 0.441%, mannitolof about 4.413% and polyacrylic acid of about 0.882% were added to themixture to obtain a suspension. The suspension was further allowed toswell by stirring. To this suspension, effective amount of glycerol ofabout 2.206% was added. The pH was maintained at about 6.3 to 6.4 withthe addition of effective amount of triethanolamine. Finallytherapeutically effective amount of hyaluronic acid of about 3.530%,followed by therapeutically effective amount of the epidermal growthfactor (EGF) of about 0.079% was added to obtain the composition fortreating the wound.

Example 8

A mixture was obtained by dissolving about 0.165% of sodium methylparaben and about 0.018% of sodium propyl paraben in purified water ofabout 91.521%. Subsequently L-lysine hydrochloride of about 0.458%,mannitol of about 4.576% and polyacrylic acid of about 0.915% were addedto the mixture to obtain a suspension. The suspension was furtherallowed to swell by stirring. To this suspension effective amount ofglycerol of about 2.288% was added. The pH was maintained at about 6.3to 6.4 with the addition of effective amount of triethanolamine. Finallytherapeutically effective amount of epidermal growth factor (EGF) ofabout 0.014% followed by Lysostaphin in therapeutically effective amountof about 0.046& was added to obtain the composition for treating thewound.

Example 9

A mixture was obtained by dissolving about 0.164% of sodium methylparaben and about 0.018% of sodium propyl paraben in purified water ofabout 91.424%. Subsequently L-lysine hydrochloride of about 0.457%,mannitol of about 4.570% and polyacrylic acid of about 0.914% were addedto the mixture to obtain a suspension. The suspension was furtherallowed to swell by stirring. To this suspension effective amount ofglycerol of about 2.285% was added. The pH was maintained at about 6.3to 6.4 with the addition of effective amount of triethanolamine.Finally, therapeutically effective amount of epidermal growth factor(EGF) of about 0.082% followed by Lysostaphin in therapeuticallyeffective amount of about 0.082% was added to obtain the composition fortreating the wound.

Example 10

A mixture was obtained by dissolving about 0.162 of sodium methylparaben and 0.018% of sodium propyl paraben in purified water of about90.232%. Subsequently, about 0.451% of L-lysine hydrochloride, about4.512% of mannitol and 0.902% of polyacrylic acid in were added to themixture to obtain a suspension. The suspension was further allowed toswell by stirring. To this suspension effective amount of glycerol ofabout 2.256% was added. The pH was maintained at about 6.3 to 6.4 withthe addition of effective amount of triethanolamine. Finally,therapeutically effective amount of epidermal growth factor (EGF) ofabout 0.005%, followed by effective amounts of aloe vera of about0.902%, vitamin E of about 0.541% and vitamin C of about 0.018% wereadded to obtain the composition for treating the wound.

Example 11

A mixture was obtained by dissolving about 0.154% of sodium methylparaben and 0.017% of sodium propyl paraben in purified water of about85.752%. Subsequently, about 0.429% of L-lysine hydrochloride, about4.288% of mannitol and 0.858% of polyacrylic acid in were added to themixture to obtain a suspension. The suspension was further allowed toswell by stirring. To this suspension effective amount of glycerol ofabout 2.144% was added. The pH was maintained at about 6.3 to 6.4 withthe addition of effective amount of triethanolamine. Finally,therapeutically effective amount of epidermal growth factor (EGF) ofabout 0.013%, followed by effective amounts of aloe vera of about5.145%, vitamin E of about 0.858% and vitamin C of about 0.343% wereadded to obtain the composition for treating the wound.

Example 12

A mixture was obtained by dissolving about 0.147% of sodium methylparaben and 0.016% of sodium propyl paraben in purified water of about81.907%. Subsequently, about 0.410% of L-lysine hydrochloride, about4.095% of mannitol and 0.819% of polyacrylic acid in were added to themixture to obtain a suspension. The suspension was further allowed toswell by stirring. To this suspension effective amount of glycerol ofabout 2.048% was added. The pH was maintained at about 6.3 to 6.4 withthe addition of effective amount of triethanolamine. Finallytherapeutically effective amount of epidermal growth factor (EGF) ofabout 0.074%, followed by effective amounts of aloe vera of about8.191%, vitamin E of about 1.474% and vitamin C of about 0.819% wereadded to obtain the composition for treating the wound.

Example 13

A mixture was obtained by dissolving about 0.149% of sodium methylparaben and about 0.017% of sodium propyl paraben in purified water ofabout 82.757%. Subsequently about 0.414% of L-lysine hydrochloride,about 4.138% of mannitol and about 0.828% of polyacrylic acid ineffective amounts were added to the mixture to obtain a suspension. Thesuspension was further allowed to swell by stirring. To this suspensioneffective amount of glycerol of about 2.069% was added. The pH wasmaintained at about 6.3 to 6.4 with the addition of effective amount oftriethanolamine. Finally, therapeutically effective amount of epidermalgrowth factor (EGF) of 0.005%, followed by effective amounts of aloevera of about 0.828%, turmeric of about 8.276%, sandal wood of about0.248% and honey of about 0.273% were added to obtain the compositionfor treating the wound.

Example 14

A mixture was obtained by dissolving about 0.098% of sodium methylparaben and about 0.011% of sodium propyl paraben in purified water ofabout 54.478%. Subsequently about 0.273% of L-lysine hydrochloride,about 2.734% of mannitol and about 0.547% of polyacrylic acid ineffective amounts were added to the mixture to obtain a suspension. Thesuspension was further allowed to swell by stirring. To this suspensioneffective amount of glycerol of about 1.367% was added. The pH wasmaintained at about 6.3 to 6.4 with the addition of effective amount oftriethanolamine. Finally, therapeutically effective amount of epidermalgrowth factor (EGF) of about 0.049%, followed by effective amounts ofaloe vera of about 0.984%, turmeric of about 38.274%, sandal wood ofabout 0.492% and honey of about 0.492% were added to obtain thecomposition for treating the wound.

Example 15

A mixture was obtained by dissolving about 0.164% of sodium methylparaben and about 0.018% of sodium propyl paraben in purified water ofabout 91.553%. Subsequently L-lysine hydrochloride of about 0.457%,mannitol of about 4.577% and polyacrylic acid of about 0.915% in wereadded to the mixture to obtain a suspension. The suspension was furtherallowed to swell by stirring. To this suspension effective amount ofglycerol of about 2.288% was added. The pH was maintained at about 6.3to 6.4 with the addition of effective amount of triethanolamine.Finally, therapeutically effective amount of epidermal growth factor(EGF) about 0.010% followed by effective amount of the protein freeblood extract of about 0.018% were added to obtain the composition fortreating the wound.

Example 16

A mixture was obtained by dissolving about 0.165% of sodium methylparaben and about 0.018% of sodium propyl paraben in purified water ofabout 91.483%. Subsequently L-lysine hydrochloride of about 0.457%,mannitol of about 4.574% and polyacrylic acid of about 0.915% in wereadded to the mixture to obtain a suspension. The suspension was furtherallowed to swell by stirring. To this suspension effective amount ofglycerol of about 2.287% was added. The pH was maintained at about 6.3to 6.4 with the addition of effective amount of triethanolamine.Finally, therapeutically effective amount of epidermal growth factor(EGF) about 0.082% followed by effective amount of the protein freeblood extract of about 0.018% were added to obtain the composition fortreating the wound.

Example 17

A mixture was obtained by dissolving about 0.165% of sodium methylparaben and 0.018% of sodium propyl paraben in purified water of about91.560%. Subsequently L-lysine hydrochloride of about 0.458%, mannitolof about 4.578% and polyacrylic acid of about 0.916% were added to themixture to obtain a suspension. The suspension was further allowed toswell by stirring. To this suspension effective amount of glycerol ofabout 2.289% was added. The pH was maintained at about 6.3 to 6.4 withthe addition of effective amount of triethanolamine. Finally,therapeutically effective amount of epidermal growth factor (EGF) ofabout 0.014% followed by effective amount of GM-CSF of about 0.003% wereadded to obtain the composition for treating the wound.

Example 18

A mixture was obtained by dissolving about 0.165% of sodium methylparaben and 0.018% of sodium propyl paraben in purified water of about91.496%. Subsequently L-lysine hydrochloride of about 0.457%, mannitolof about 4.574% and polyacrylic acid of about 0.914% were added to themixture to obtain a suspension. The suspension was further allowed toswell by stirring. To this suspension effective amount of glycerol ofabout 2.287% was added. The pH was maintained at about 6.3 to 6.4 withthe addition of effective amount of triethanolamine. Finally,therapeutically effective amount of epidermal growth factor (EGF) ofabout 0.082% followed by effective amount of GM-CSF of about 0.003% wereadded to obtain the composition for treating the wound.

Example 19

A mixture was obtained by dissolving about 0.164% of sodium methylparaben and about 0.018% of sodium propyl paraben in purified water ofabout 90.567%. Subsequently L-lysine hydrochloride of about 0.460%,mannitol of about 4.550% and polyacrylic acid of about 0.910% were addedto the mixture to obtain a suspension. The suspension was furtherallowed to swell by stirring. To this suspension effective amount ofglycerol of about 2.270% was added. The pH was maintained at about 6.3to 6.4 with the addition of effective amount of triethanolamine.Finally, therapeutically effective amount of epidermal growth factor(EGF) of about 0.010% followed by effective amounts of Beta-1,3-D-glucanof about 0.550%, were added to obtain the composition for treating thewound.

Example 20

A mixture was obtained by dissolving about 0.163% of sodium methylparaben and about 0.018% of sodium propyl paraben in purified water ofabout 90.567%. Subsequently L-lysine hydrochloride of about 0.453%,mannitol of about 4.528% and polyacrylic acid of about 0.906% were addedto the mixture to obtain a suspension. The suspension was furtherallowed to swell by stirring. To this suspension effective amount ofglycerol of about 2.264% was added. The pH was maintained at about 6.3to 6.4 with the addition of effective amount of triethanolamine.Finally, therapeutically effective amount of epidermal growth factor(EGF) of about 0.014% followed by effective amounts of Beta-1,3-D-glucanof about 1.087%, were added to obtain the composition for treating thewound.

Example 21

A mixture was obtained by dissolving about 0.165% of sodium methylparaben and about 0.018% sodium propyl paraben in purified water ofabout 91.565%. Subsequently L-lysine hydrochloride of about 0.458%,mannitol of about 4.578% and polyacrylic acid of about 0.916% were addedto the mixture to obtain a suspension. The suspension was furtherallowed to swell by stirring. To this suspension effective amount ofglycerol of about 2.289% was added. The pH was maintained at about 6.3to 6.4 with the addition of effective amount of triethanolamine.Finally, therapeutically effective amount of epidermal growth factor(EGF) of about 0.005% followed by effective amount of PDGF of about0.005% were added to obtain the composition for treating the wound.

Example 22

A mixture was obtained by dissolving about 0.165% of sodium methylparaben and about 0.018% sodium propyl paraben in purified water of91.424%. Subsequently L-lysine hydrochloride of about 0.457%, mannitolof about 4.571% and polyacrylic acid of about 0.914% were added to themixture to obtain a suspension. The suspension was further allowed toswell by stirring. To this suspension effective amount of glycerol ofabout 2.286 was added. The pH was maintained at about 6.3 to 6.4 withthe addition of effective amount of triethanolamine. Finally,therapeutically effective amount of epidermal growth factor (EGF) 0.082%followed by effective amount of PDGF of about 0.082% were added toobtain the composition for treating the wound.

Example 23

A mixture was obtained by dissolving about 0.164% of sodium methylparaben and about 0.018% of sodium propyl paraben in purified water ofabout 91.286%. Subsequently L-lysine hydrochloride of about 0.456%,mannitol of about 4.564%, polyacrylic acid of about 0.913%, zinc oxideof about 0.018%, salicyclic acid of about 0.018%, clobetasol propioateof about 0.274% were added to the mixture to obtain a suspension. Thesuspension was further allowed to swell by stirring. To this suspensioneffective amount of glycerol of about 2.282% was added. The pH wasmaintained at about 6.3 to 6.4 with the addition of effective amount oftriethanolamine. Finally, a therapeutically effective amount ofepidermal growth factor (EGF) of about 0.005% was added to obtain thecomposition for treating the wound.

Example 24

A mixture was obtained by dissolving about 0.157% of sodium methylparaben and about 0.017% of sodium propyl paraben in purified water ofabout 87.478%. Subsequently L-lysine hydrochloride of about 0.437%,mannitol of about 4.374%, polyacrylic acid of about 0.876%, zinc oxideof about 2.187%, salicyclic acid of about 1.750%, clobetasol propionateof about 0.525% were added to the mixture to obtain a suspension. Thesuspension was further allowed to swell by stirring. To this suspension,effective amount of glycerol of about 2.187% was added. The pH wasmaintained at about 6.3 to 6.4 with the addition of effective amount oftriethanolamine. Finally, a therapeutically effective amount ofepidermal growth factor (EGF) of about 0.013% was added to obtain thecomposition for treating the wound.

Example 25

A mixture was obtained by dissolving about 0.152% of sodium methylparaben and about 0.017% of sodium propyl paraben in of purified waterof about 84.610. Subsequently L-lysine hydrochloride of about 0.423%,mannitol of about 4.230%, polyacrylic acid of about 0.846%, zinc oxideof about 3.384%, salicyclic acid of about 3.384%, clobetasol propionateof about 0.761% were added to the mixture to obtain a suspension. Thesuspension was further allowed to swell by stirring. To this suspensioneffective amount of glycerol of about 2.115% was added. The pH wasmaintained at about 6.3 to 6.4 with the addition of effective amount oftriethanolamine. Finally, a therapeutically effective amount ofepidermal growth factor (EGF) of about 0.076% was added to obtain thecomposition for treating the wound.

Example 26

A mixture was obtained by dissolving about 0.164% of sodium methylparaben and about 0.018% of sodium propyl paraben in of purified waterof about 91.490%. Subsequently L-lysine hydrochloride of about 0.457%,about 4.570% of mannitol and about 0.914% of polyacrylic acid were addedto the mixture to obtain a suspension. The suspension was furtherallowed to swell by stirring. To this suspension effective amount ofglycerol of about 2.287% was added. The pH was maintained at about 6.3to 6.4 with the addition of triethanolamine. Finally, therapeuticallyeffective amounts of epidermal growth factor (EGF) of about 0.010%followed by effective amount of silver sulphadiazine of about 0.091%were added to obtain the composition for treating the wound.

Example 27

A mixture was obtained by dissolving about 0.16% of sodium methylparaben and about 0.020% of sodium propyl paraben in of purified waterof about 91.360%. Subsequently L-lysine hydrochloride of about 0.46%,about 4.570% of mannitol and about 0.910% of polyacrylic acid were addedto the mixture to obtain a suspension. The suspension was furtherallowed to swell by stirring. To this suspension effective amount ofglycerol of about 2.280% was added. The pH was maintained at about 6.3to 6.4 with the addition of triethanolamine. Finally, therapeuticallyeffective amount of epidermal growth factor (EGF) of about 0.010%followed by effective amount of silver sulphadiazine of about 0.230%were added to obtain the composition for treating the wound.

Example 28

A mixture was obtained by dissolving about 0.164% of sodium methylparaben and about 0.018% of sodium propyl paraben in purified water ofabout 91.228%. Subsequently L-lysine hydrochloride of about 0.456%,about 4.561% of mannitol and about 0.912% of polyacrylic acid were addedto the mixture to obtain a suspension. The suspension was furtherallowed to swell by stirring. To this suspension effective amount ofglycerol of about 2.281% was added. The pH was maintained at about 6.3to 6.4 with the addition of triethanolamine. Finally, therapeuticallyeffective amount of epidermal growth factor (EGF) of about 0.014%followed by effective amount of silver sulphadiazine of about 0.365%were added to obtain the composition for treating the wound.

Example 29

A mixture was obtained by dissolving about 0.155% of sodium methylparaben and about 0.017% of sodium propyl paraben in purified water ofabout 85.873%. Subsequently L-lysine hydrochloride of about 0.429%,mannitol of about 4.294% and polyacrylic acid of about 0.859% were addedto the mixture to obtain a suspension. The suspension was furtherallowed to swell by stirring. To this suspension effective amount ofglycerol of about 2.147% was added. The pH was maintained at about 6.3to 6.4 with the addition of triethanolamine. Finally therapeuticallyeffective amounts of epidermal growth factor (EGF) of about 0.001%,followed by effective amounts of hydrocortisone acetate of about 0.215%,lidocaine of about 2.147%, zinc oxide of about 3.435%, and allantoin ofabout 0.429% to obtain the composition for treating the wound.

Example 30

A mixture was obtained by dissolving about 0.155% of sodium methylparaben and about 0.017% of sodium propyl paraben in purified water ofabout 85.859%. Subsequently L-lysine hydrochloride of about 0.429%,mannitol of about 4.293% and polyacrylic acid of about 0.859% were addedto the mixture to obtain a suspension. The suspension was furtherallowed to swell by stirring. To this suspension effective amount ofglycerol of about 2.147% was added. The pH was maintained at about 6.3to 6.4 with the addition of triethanolamine. Finally, therapeuticallyeffective amount of epidermal growth factor (EGF) of about 0.005%,followed by effective amounts of hydrocortisone acetate of about 0.215%,lidocaine of about 2.147%, zinc oxide of about 3.435%, and allantoin ofabout 0.429% to obtain the composition for treating the wound.

Example 31

A mixture was obtained by dissolving about 0.151% of sodium methylparaben and about 0.017% of sodium propyl paraben in purified water ofabout 83.742%. Subsequently L-lysine hydrochloride of about 0.419%,mannitol of about 4.187% and polyacrylic acid of about 0.837% were addedto the mixture to obtain a suspension. The suspension was furtherallowed to swell by stirring. To this suspension effective amount ofglycerol of about 2.094% was added. The pH was maintained at about 6.3to 6.4 with the addition of triethanolamine. Finally therapeuticallyeffective amounts of epidermal growth factor (EGF) of about 0.013%,followed by effective amounts of hydrocortisone acetate of about 0.167%,lidocaine of about 2.931%, zinc oxide of about 4.187%, and allantoin ofabout 1.256% to obtain the composition for treating the wound.

Example 32

A mixture was obtained by dissolving about 0.179% of sodium methylparaben and about 0.020% of sodium propyl paraben in purified water ofabout 99.034%. Subsequently L-lysine hydrochloride of about 0.419%, andpolyacrylic acid in effective amount of about 0.249% was added to themixture to obtain a suspension. The suspension was further allowed toswell by stirring. The pH was maintained at about 6.3 to 6.4 with theaddition of triethanolamine. Finally, a therapeutically effective amountof epidermal growth factor (EGF) in the range of about 0.099% was addedto obtain the composition for treating the wound.

Example 33

A mixture was obtained by dissolving about 0.177% of sodium methylparaben and about 0.020% of sodium propyl paraben in purified water ofabout 97.765%. Subsequently, L-lysine hydrochloride of about 0.419%, andpolyacrylic acid in effective amount of about 0.736% was added to themixture to obtain a suspension. The suspension was further allowed toswell by stirring. The pH was maintained at about 6.3 to 6.4 with theaddition of triethanolamine. Finally, a therapeutically effective amountof epidermal growth factor (EGF) in the range of about 0.884% was addedto obtain the composition for treating the wound.

Example 34

An effective amount of purified water of about 98.131% was initiallyheated to 80 degree C. To this water, effective amount ofcarboxymethylcellulose of about 1% was added and the mixture was allowedto swell by stirring. The mixture obtained was sterilized at 121 degreeC. for 15 minutes. The sterilized mixture was then allowed to cool.Subsequently L-lysine hydrochloride of about 0.419% was added. Finally,therapeutically effective amount of epidermal growth factor (EGF) ofabout 0.45% was added and allowed to mix thoroughly to obtain thecomposition for treating the wound.

Example 35

An effective amount of purified water of about 97.381% was initiallyheated to 80 degree C. To this water, effective amount ofcarboxymethylcellulose of about 1.64% was added and the mixture wasallowed to swell by stirring. The mixture obtained was sterilized at 121degree C. for 15 minutes. The sterilized mixture was then allowed tocool. Subsequently L-lysine hydrochloride of about 0.419% was added.Finally, therapeutically effective amounts of epidermal growth factor(EGF) of about 0.9% was added and allowed to mix thoroughly to obtainthe composition for treating the wound.

The process of addition of the ingredients can be carried out in thesame sequence or in different sequences, which can give the same stableformulation known to those experienced in the art of formulations. Thescope of the invention is not limited to the addition of same excipientsor other excipients having similar functions or different concentrationsin the same sequence but also to other addition sequences which can givestable formulation or similar effects.

All the compositions were tested for the EGF activity by ELISA and allthe other ingredients were tested by the pharmacopeial methods or otherappropriately validated methods which clearly confirm the activity.

EGF activity of the compositions was tested by means of proliferation ofthe 3T3 cell lines. Briefly, in this method, the known quantity of 3T3cell were incubated with different quantities of standard EGF andformulated EGF and incubated at 37 degree C. for 5-8 days. Cell countwas estimated and it was compared with the controls.

Results confirmed that the proliferation of cells treated with EGF wassignificantly higher as compared to the controls. Similarly SDS profilealso showed the maintenance of the same profile for the entire period ofstudy.

The scope of the combination is not limited just by the above range, butalso to other combinations, which can still maintain the same functionalactivity of EGF known to those who are skilled in the art.

In the foregoing specification, specific embodiments of the presentinvention have been described. However, one of ordinary skill in the artappreciates that various modifications and changes can be made withoutdeparting from the scope of the present invention as set forth in theclaims below. Accordingly, the specification and the examples are to beregarded in an illustrative rather than a restrictive sense, and allsuch modifications are intended to be included within the scope ofpresent invention. The benefits, advantages, solutions to problems, andany element(s) that may cause any benefit, advantage, or solution tooccur or become more pronounced are not to be construed as a critical,required, or essential features or elements of any or all the claims.The invention is defined solely by the appended claims including anyamendments made during the dependency of this application and allequivalents of those claims as issued.

What is claimed:
 1. A composition comprising recombinant human epidermalgrowth factor present in a range of about 0.001% to about 0.9% (w/w);L-lysine hydrochloric acid present in the range of about 0.2% to about0.5% (w/w) and mannitol present in the range of about 1% to about 10%(w/w) as stabilizers; sodium methyl paraben present in the range ofabout 0.016% to about 0.18% (w/w) and sodium propyl paraben present inthe range of about 0.01% to about 0.02% (w/w) as preservatives; polyacrylic acid present in the range of about 0.25% to about 1% (w/w) as athickening agent; water present in the range of about 50% to about 99.5%(w/v) as a carrier; triethanol amine present in an effective amount tomaintain the pH of the composition between 6.3 to 6.4 as a pH regulatingagent; glycerol present in the range of about 1% to about 2.5% (w/w) asa humectant; and a therapeutic agent present in a range of about 0.005%to about 0.5% (w/w).
 2. The composition of claim 1, wherein therecombinant human epidermal growth factor is in a topical formulation inthe form of gel, spray, ointment, cream and lotion, in oral formulationin the form of tablet and capsule, or in parenteral formulation in theform of injections.
 3. The composition of claim 1, wherein thetherapeutic agent is an antiinfective agent silver sulphadiazine presentin a range of about 0.005% to about 0.5% (w/w).
 4. The composition ofclaim 1, wherein the therapeutic agent is a recombinant antibioticlysostaphin present in the range of about 0.01% to about 0.1% (w/w). 5.The composition of claim 1, wherein the therapeutic agent is hyaluronicacid present in the range of about 0.05% to about 5% (w/w).
 6. Thecomposition of claim 1, wherein the therapeutic agent is GranulocyteMacrophage Colony Stimulating Factor (GM-CSF) present in the range ofabout 0.001% to 0.005% (w/w).
 7. The composition of claim 1, wherein thetherapeutic agent is Platelet Derived Growth Factor present in the rangeof about 0.005% to about 0.01% (w/w).
 8. The composition of claim 1,wherein the therapeutic agent is an immune modulator beta 1,3-D-glucanpresent in the range of about 0.5% to about 2% (w/w).
 9. The compositionof claim 1, wherein the therapeutic agent is present in a combinationwith a vaso constrictor hydrocortisone acetate present in the range ofabout 0.215% (w/w), zinc oxide present in the range of about 0.015% toabout 5% (w/w), lindocaine present in the range of about 2% to 3% (w/w),and allantoin present in the range of about 0.4% to about 1.5% (w/w).10. The composition of claim 1, wherein the therapeutic agent is presentin a combination of natural product(s) consisting of aloevera; turmeric;sandalwood; and honey.
 11. The composition of claim 1, wherein thetherapeutic agent is present in a combination of natural product(s)consisting of aloevera; Vitamin E and Vitamin C.
 12. The composition ofclaim 1, wherein the therapeutic agent is protein free blood extract,present in the range of about 0.015% to 0.02% (w/w).
 13. The compositionof claim 1, wherein the therapeutic agent is present in combination ofzinc oxide present in the range of about 0.015% to about 5% (w/w),salicylic acid present in the range of about 1% to about 5% (w/w) andclobetasol propionate present in the range of about 0.1% to about 1%(w/w).